The DUBscan™ kit has been flexibly designed for many potential applications, including the identification of DUB enzymes which support the deubiquitylation of substrate proteins.
The kit contains a DUBscan plate (a panel of DUBs arrayed across 47 wells of a 96 well plate) plus a control ubiquitylation reaction mix for performing both positive and negative control deubiquitylation reactions. Through the addition of your own DUB substrate of interest to the wells of a DUBscan plate you can screen the DUB enzyme panel in about one hour. Reaction products may be analyzed as required, for example by SDS-PAGE and Western blotting using specific antibodies. One kit contains all the reagents and buffers necessary to perform a DUBscan of the panel of 47 DUB assay wells.
Utility of the DUBscan Kit
- Examine the cleavage of in vitro ubiquitylated substrates generated using different E2/E3 combinations
- Examine the cleavage of in vivo ubiquitylated substrates after capture from a cell lysate
- Investigate DUB ubiquitin-ubiquitin linkage cleavage specificity
- Examine the relative position of cleavage within ubiquitin chains (distal versus proximal)
- Explore mono and poly-deubiquitylation DUB specificity
- Explore potential DUB inhibitors and/or activators
- Identify novel DUB binding proteins
- Explore how DUBs may interact with and modify the activity of other ubiquitin system proteins such as E2s